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Title: Ribulose diphosphate carboxylase of the cyanobacterium Spirulina platensis

Abstract

The ribulose diphosphate (RDP) carboxylase activity of the cyanobacterium Spirulina platensis is represented by two peaks when a cell homogenate is centrifuged in a sucrose density gradient. In the case of differential centrifugation (40,000 g, 1 h), the activity of the enzyme was distributed between the supernatant liquid (soluble form) and the precipitate (carboxysomal form). From the soluble fraction, in which 80-95% of the total activity of the enzyme is concentrated, electrophoretically homogeneous RDP carboxylase was isolated by precipitation with ammonium sulfate and centrifugation in a sucrose density gradient. The purified enzyme possessed greater electrophoretic mobility in comparison with the RDP carboxylase of beans Vicia faba. The molecular weight of the enzyme, determined by gel filtration, was 450,000. The enzyme consists of monotypic subunits with a molecular weight of 53,000. The small subunits were not detected in electrophoresis in polyacrylamide gel in the presence of SDS after fixation and staining of the gels by various methods.

Authors:
; ;
Publication Date:
Research Org.:
A.N. Bakh Institute of Biochemistry, Moscow, USSR
OSTI Identifier:
5856545
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemistry (Engl. Transl.); (United States); Journal Volume: 51:5; Other Information: Translated from Biokhimiya; 51: No. 5, 816-822(May 1986)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; CARBOXY-LYASES; ELECTROPHORESIS; ENZYME ACTIVITY; RADIOENZYMATIC ASSAY; CYANOBACTERIA; CARBON 14 COMPOUNDS; DIALYSIS; METABOLISM; MOLECULAR WEIGHT; ORGANIC PHOSPHORUS COMPOUNDS; RIBULOSE; UPTAKE; CARBOHYDRATES; CARBON-CARBON LYASES; ENZYMES; KETONES; LABELLED COMPOUNDS; LYASES; MICROORGANISMS; MONOSACCHARIDES; ORGANIC COMPOUNDS; PENTOSES; SACCHARIDES; SEPARATION PROCESSES; 550201* - Biochemistry- Tracer Techniques

Citation Formats

Terekhova, I.V., Chernyad'ev, I.I., and Doman, N.G.. Ribulose diphosphate carboxylase of the cyanobacterium Spirulina platensis. United States: N. p., 1986. Web.
Terekhova, I.V., Chernyad'ev, I.I., & Doman, N.G.. Ribulose diphosphate carboxylase of the cyanobacterium Spirulina platensis. United States.
Terekhova, I.V., Chernyad'ev, I.I., and Doman, N.G.. Thu . "Ribulose diphosphate carboxylase of the cyanobacterium Spirulina platensis". United States. doi:.
@article{osti_5856545,
title = {Ribulose diphosphate carboxylase of the cyanobacterium Spirulina platensis},
author = {Terekhova, I.V. and Chernyad'ev, I.I. and Doman, N.G.},
abstractNote = {The ribulose diphosphate (RDP) carboxylase activity of the cyanobacterium Spirulina platensis is represented by two peaks when a cell homogenate is centrifuged in a sucrose density gradient. In the case of differential centrifugation (40,000 g, 1 h), the activity of the enzyme was distributed between the supernatant liquid (soluble form) and the precipitate (carboxysomal form). From the soluble fraction, in which 80-95% of the total activity of the enzyme is concentrated, electrophoretically homogeneous RDP carboxylase was isolated by precipitation with ammonium sulfate and centrifugation in a sucrose density gradient. The purified enzyme possessed greater electrophoretic mobility in comparison with the RDP carboxylase of beans Vicia faba. The molecular weight of the enzyme, determined by gel filtration, was 450,000. The enzyme consists of monotypic subunits with a molecular weight of 53,000. The small subunits were not detected in electrophoresis in polyacrylamide gel in the presence of SDS after fixation and staining of the gels by various methods.},
doi = {},
journal = {Biochemistry (Engl. Transl.); (United States)},
number = ,
volume = 51:5,
place = {United States},
year = {Thu Nov 20 00:00:00 EST 1986},
month = {Thu Nov 20 00:00:00 EST 1986}
}
  • The photosynthetic performance of a helical tubular photobioreactor (``Biocoil``), incorporating the filamentous cyanobacterium Spirulina platensis, was investigated. The photobioreactor was constructed in a cylindrical shape with a 0.25-m{sup 2} basal area and a photostage comprising 60 m of transparent PVC tubing of 1.6-cm inner diameter. The inner surface of the cylinder was illuminated with cool white fluorescent lamps; the energy input of photosynthetically active radiation into the photobioreactor was 2,920 kJ per day. An air-lift system incorporating 4% CO{sub 2} was used to circulate the growth medium in the tubing. The maximum productivity achieved in batch culture was 7.18 gmore » dry biomass per day which corresponded to a photosynthetic (PAR) efficiency of 5.45%. The CO{sub 2} was efficiently removed from the gaseous stream; monitoring the CO{sub 2} in the outlet and inlet gas streams showed a 70% removal of CO{sub 2} from the inlet gas over an 8-h period with almost maximum growth rate.« less
  • In the presence of D-ribulose diphosphate, crystalline ribulose diphosphate carboxylase from Nicotiana tabacum leaves undergoes a profound change in solubility. The solubility change did not involve a conformational change in molecular volume exceeding 2 percent as measured by sedimentation velocity suggesting no gross change in quaternary structure. However, the change in solubility did involve a tertiary structural change wherein some previously buried tyrosyl and tryptophyl residues became exposed, as indicated by difference spectrophotometry. Although the enzyme molecule has 8 binding sites for ribulose diphosphate, the conformational change is complete after 4 substrate molecules are bound. A cooperative action among themore » subunits is proposed.« less
  • Ribulose 1,5-diphosphate carboxylase, when activated by preincubation with 10 mM MgCl/sub 2/ and 1 mM bicarbonate in the absence of ribulose 1,5-diphosphate, can be further activated about 170% with 0.5 mM NADPH present in the preincubation mixture. NADP/sup +/, NADH, and NAD/sup +/ are ineffective. The activation by NADPH is comparable to that previously seen with 0.05 to 0.10 mM 6-phosphogluconate in that these specific preincubation conditions are required, but the effects of NADPH and 6-phosphogluconate is not additive. Moreover, where higher concentrations of 6-phosphogluconate inhibited the enzyme, higher concentrations of NADPH give a greater activation, saturating at about 1more » mM and 200%. Under the specified conditions of preincubation, fructose 1,6-diphosphate has an activation curve similar to that of 6-phosphogluconate, peaking at 0.1 mM and 70%. Above this level activation decreases, and inhibition is seen at still higher concentrations. Other metabolites tested produced smaller or no effects on the enzyme activity assayed under these conditions. When either reduced NADP or 6-phosphogluconate is present in the preincubation mixture, it becomes possible to determine the Km for bicarbonate using a Lineweaver--Burk plot, and the Km for bicarbonate under these conditions is 2.8 mM, corresponding to 0.3% CO/sub 2/ at pH 7.8 and 25 C.« less